This application is submitted in response to PAR-13-095 with the intent of testing lineage committed cells derived from human induced pluripotent stem cells (hiPSC) for the treatment of congenital lung diseases. The approaches outlined in this proposal build on the expertise of our team in developmental ontogeny, tissue engineering, stem/progenitor cell studies, novel in vivo imaging modalities, and a translational nonhuman primate model. We have studied methods to differentiate human pluripotent cells towards lung-related lineages including airway epithelial cells (AECs); engineered constructs with unique scaffolds and matrices; and developed techniques to effectively label, transplant, and monitor the fate of transplanted precursors in vivo in rhesus monkeys. We are now poised to advance our prior discoveries and break new ground to address key questions related to iPSC-derived precursors to treat congenital lung diseases, such as pulmonary surfactant deficiency, through the following Specific Aims: (1) Optimize lineage commitment of hiPSCs in 3D culture to enhance AEC differentiation and purity, and (2) Conduct proof-of-concept studies to evaluate fetal transplant efficiency and cell fate in vivo. We will optimize proliferation and integration o key cell populations in 3D culture to enhance lineage specification, and evaluate transplant efficiency and cell fate in vivo through innovative imaging paradigms including bioluminescence imaging and positron emission tomography/computed tomography (PET/CT). The lung has limited capacity for repair, and regenerative concepts that are based on developmental principles, tailored to the age of the patient, tested in a translational primate model, and use state-of-the-art imaging technologies could have a major impact on the field. These investigations will provide new insights and guide new avenues of translational research that could have significant impact and compelling benefits for children and all age groups across the lifespan.